A comparative study of photoreactive tRNA derivatives containing a 6-thioguanosine or diazirine derivative of 5-methyleneaminouridine was performed using Escherichia coli ribosomes. The conditions of producing functional tRNAs containing photoreactive residues have been selected using T7 RNA polymerase transcription. Photochemical modification was conducted upon binding the corresponding tRNA Phe transcripts carrying the photoreactive residues of nucleotide analogs to the P site of the ribosome. The results of modification were analyzed using oligonucleotide-directed hydrolysis of rRNA by RNase H, as well as reverse transcription. It is shown that 6-thioguanosine is a promising agent for studying RNA-protein complexes by photochemical crosslinking methods.
|Number of pages||7|
|Publication status||Published - Sep 2004|
- Escherichia coli
- photoaffinity modification